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MarkW wrote:Hello Guys,
I read this as theory not known facts. Does the gene CYP27B1 do this is all pwMS? To what degree? Is there evidence? Does D3 from sunlight need this gene? It would be great if a pulse of calcitrol then D3 reboots the MS immune system and keeps it operating normally. If the dose for a 70kg human (me) was a reasonable risk I would try this in the coming weeks. I need to study the paper and supporting papers in detail. We need to keep in mind that there is a D3 blood level/disease activity level relationship so there are more variables to consider. Like Ed says the costs of taking calcitrol/D3 is very low so funding research will be a problem.
kind regards,
Markw

Mark,

While I appreciate what you are saying, I think it is fair to characterize Prof. Hayes' findings as facts, but facts limited at this point to EAE mice. She has conducted a series of three major studies of this leading to her latest paper which brings this all together.

CYP27B1 is the gene that expresses the protein that converts D3 to calcitriol in people, pwMS and mice. In pwMS, this gene is unable to do this in the CNS at the site of the inflammatory process associated with MS. The gene functions elsewhere in the body or we would have no calcitriol and MS would be the least of your worries. :>)

As far as EAE and considerable research related to pwMS, there is a lot of evidence and measurement of the effect of this in terms of the proliferation of CD4+ T-cells and the fact that a single, high dose of calcitriol is sufficient to set off apoptosis which kills them off. That, along with other beneficial effects of the calcitriol is enough to effectively reboot the immune system in the CNS. It stops the disease process and keeps it in remission.

The studies are conclusive in this regard. What is not known is whether this will work the same way in humans.

I have copies of a group of relevant papers which I can send anyone who doesn't have free access. PM me with an email addy and I'll send them to you. One is Prof Hayes Calcitriol + D3 study. The second is a study by the Ebers group on the long term use of calcitriol in pwMS. The final paper is a dose escalation study of calcitriol in pwCancer which may or may not be relevant to pwMS.

Based on calculation of the human equivalent of the calcitriol doses being given to the EAE mice, the proposed starting dose to test whether this will work in humans is 70% as much as the Maximum Tolerated Dose in pwCancer.

I have used calcitriol in cats for over 13 years and wouldn't hesitate to take a single dose of calcitriol at this level just to test the effects. And if I had MS, I wouldn't hesitate for a nanosecond to see if this works.

I am in discussion with 6 medical professionals who have MS to see if they will pioneer the use of calcitriol + D3 in humans to assess the safety and see if it moves the needle.

And I have discussed getting a formal clinical trial organized and funded to test this in a small, IRB-approved trial to again, see if it is safe and moves the needle on MS.

As these efforts become more finalized, I will keep everyone posted on them here.

Wanted to clarify something related to my previous message.

I said I would not hesitate to take the dose of calcitriol needed to see if it is safe and would do it in a heartbeat if I had MS.

But, I think we all need to wait until a small group of people who by training are better able to judge the risks involved, ie, medical professionals with MS, test it and then we do at least one Phase 1 formal, IRB-approved clinical trial to be sure this is safe. That trial is going to need lots of financial support so we can all be a part of this. :>)

After that, I think we publish a treatment protocol that pwMS can take to their GPs (or PCPs) and try it and support any projects aimed at further large scale clinical trials.

This is a case where no one needs permission from their neurologist. Calcitriol is a shipping compound that any physician can prescribe.

Unlike CCSVI, there is NOTHING that the pharmaceutical companies or supporting neurologists can do to block this becoming widely used if testing shows both safety and effectiveness.

Unlike what happened with CCSVI, I hope we can create a patient-driven, worldwide registry to gather data on safety and effectiveness so that we don't lose valuable information like this which could help us fully understand whether this is going to help.

Duplicate Post

Squeakycat wrote:.... I think it is fair to characterize Prof. Hayes' findings as facts, but facts limited at this point to EAE mice. She has conducted a series of three major studies of this leading to her latest paper which brings this all together.
CYP27B1 is the gene that expresses the protein that converts D3 to calcitriol in people, pwMS and mice. In pwMS, this gene is unable to do this in the CNS at the site of the inflammatory process associated with MS. The gene functions elsewhere in the body or we would have no calcitriol and MS would be the least of your worries. :>)
................I will keep everyone posted on them here.

Hello Ed,
Let's agree to differ on facts. I have not seen hard evidence on gene CYP27B1 and for me evidence from mice with EAE is not about MS. EAE is a massive attack to the mouse brain, whereas MS is chronic damage to the human brain.
I call myself an MS medical professional, plus Prof George Ebers works close to home for consultation on the dosage for MS. You noted that this would be 70% of dosage for pwcancer. For me it is already an approved safe dose !
I am pretty sure that vested interests will try to obstruct this. I would not speak about rebooting any genes, rather a safe therapy which appears to help pwMS. Marketing is key from the start.
Best wishes,
Markw

my e-mail is mark@walkerm.com.

MarkW wrote:Let's agree to differ on facts. I have not seen hard evidence on gene CYP27B1 and for me evidence from mice with EAE is not about MS. EAE is a massive attack to the mouse brain, whereas MS is chronic damage to the human brain.

Mark,

I've just emailed you the Hayes study so you can see the hard evidence regarding CYP27B1. This gene is of considerable interest these days. I am receiving 1-3 alerts each day about new papers focused on it.

Prof Hayes' evidence regarding what is happening with CYP27B1 is somewhat circumstantial. The problematic CD4+ T-cells in MS and EAE should undergo programmed cell death if sufficient calcitriol were available. They don't and they have an affinity for myelin which is what causes the damage. When she provides her mice with a high dose of calcitriol, these cells undergo apoptosis indicating that the problem isn't something internal to the CD4+ cells, but caused by a lack of sufficient quantities of calcitriol.

From that, she concludes that something is blocking the full effectiveness of CYP27B1 in converting 25(OH)D3 to calcitriol. She can directly measure the number of CD4+ cells and knows they are dying off. And she further knows that after this single dose of calcitriol, CYP27B1 must be working again because vitamin D3 alone is enough to keep the mice in remission.

That seems fairly conclusive to me, although the evidence is indirect.

As far as whether EAE in mice is like MS in humans. It is certainly not in terms of the origin of the disease, but, if you take MS to be a disease with many possible causes, EBV virus, CPn stealth bacteria, or turbulent blood flow, the common end result is a break down of the Blood Brain Barrier and the havoc that wrecks on the CNS as things cross over that should not, particular myelin attacking CD4+ T-cells.

EAE is induced with Mycobacterium tuberculosis which causes a breach in the BBB. Prof Hayes goes a step further and injects her mice with pertussis toxin to be sure that there is a breach in the BBB. So while not the same in terms of the original cause of the breach, EAE is a fairly good mimic of the state of the disease in which the BBB is broken and things are crossing over to the CNS that shouldn't be there.

With the CYP27B1 gene being essentially the same in humans and mice, I think there is every reason to expect that the results will be similar, although given the genetic diversity of the genes of pwMS and origins of the disease, and the relative genetic homogeneity of lab mice, there may well be some pwMS who will not benefit from this approach.

It still seems worth trying. Just imagine a world where we are treating MS with a naturally occurring compound that is the critical actor in the local, adaptive immune system and at a cost that is a tiny fraction of the cost of immune-suppressing DMDs. If this works, we will need to invest in antacids since their sales will likely skyrocket as pharmaceutical industry executives consume them by the gross.

MarkW wrote:You noted that this would be 70% of dosage for pwcancer. For me it is already an approved safe dose !

Mark,
I need to clarify this.

The human equivalent of the optimal dose given the EAE mice by Prof Hayes is about twice as high as any dose I have ever seen tested in the cancer studies.

What we are looking at as an initial test dose is 48 times less than the optimum dose used in the EAE mice. This dose is 70% of the Maximum Tolerated Dose in a study in cancer, but still quite high, 100 times higher than the approved dose for treating patients with renal failure.

I think there are a number of reasons this is safe, but it may not move the needle in terms of therapeutic effect. If that's the case, and the test subjects are willing, we will double it. That brings it close to the lowest therapeutic level in the Hayes study.

We are also going to double the D3 dose, still well below the recommended upper limit, but a dose that has been repeatedly shown to be safe in human trials including trials in pwMS as a partial way of offsetting the lower calcitriol dose we will be testing.

Here's a summary of why I think the proposed initial dose is safe and I think that this would be true, even if we have to double it to have effect:

1. Higher doses of calcitriol have been tested in dose escalation studies involving human cancer patients. Fakih found that that Maximum Tolerated Dose in terms of hypercalcemia was 30% higher than the proposed test dose in humans and the dose was being administered weekly for a long-term.

2. Moderately high doses of calcitriol administered daily for as long as 48 weeks have been found to be safe in people with MS in Wingerchuk. While the dose was incorrectly administered daily rather than pulsed weekly, the pulse dose would have been 35% as much as proposed and administered for a full year rather than a single dose.

3. The half life of calcitriol even at high doses is between 3-4 hours after which any unused calcitriol is converted to an inactive form and removed by the kidneys which would make any effect on calcium homeostasis transient.

4. While high doses of vitamin D and calcitriol stimulate the uptake of calcium, even higher doses have the opposite effect by suppressing PTH. This consideration is discussed in Beer involving a trial in which a dose approximately the same as proposed for MS was tested in cancer patients and administered weekly.

5. Further, pre-treatment testing will be done to establish that calcium, phosphorous levels and kidney function (BUN and creatinine) are within normal limits which should help to minimize the possibility of hypercalcemia and hyperphosphatemia.

6. Finally, the standard treatment for hypercalcemia is to withdraw administration of calcitriol and the study is based on administering a single dose of calcitriol so the protocol automatically withdraws calcitriol after the first dose.

PM me if you would like copies of the references cited here.

Morning all,
I have been going through this proposal for some time in my highly defective mind so what I say may be off the wall!

There is one factor that Ebers and co have often spoken about which is the difference between EAE and MS. The key point that I take from that because of my own bacterial infection woes is that the bacteria and or virus(es) used to induce EAE aka the breach of the BBB in Prof H's study are the 'item' that the immune system is hunting and killing!

If that infection and possible other infections that will cross the BBB to create the outcome of 'lesions and disability' the target for the treatment, then is the treatment anything to do with the symptoms? The outcome of symptoms 'can' be an incidental process (synergistic) that occurs because of a breached BBB, but not the pathogen itself or vice versa.

So is it the repaired immune system kill rate the makes a rapid recovery in the Prof H study or is it repair of the damage caused by a multi-pathogen infection within the BBB safe zone that has been entered or is it the repair of the BBB breaches or entry points?
Probably many more theories why the mice recover so quickly after a stunt of Calcitriol and Vit D?

At another level does it matter? The outcome is more important than the how!! The knowledge can appear later if the treatment is safe, effective and without risk!!

I think because the 'disease' MS has so many forms that the calcitriol and Vit D research and study will be open for hypothesis expression! ;)

So a safety test in humans is probably the step to take, paedophiles and murderers ?


Nigel

NZer1 wrote:There is one factor that Ebers and co have often spoken about which is the difference between EAE and MS. The key point that I take from that because of my own bacterial infection woes is that the bacteria and or virus(es) used to induce EAE aka the breach of the BBB in Prof H's study are the 'item' that the immune system is hunting and killing!

Nigel,
The immune system has two jobs here:

• 1. Destroying pathogens, and

• 2. Repairing damaged cells in the BBB caused by the pathogens, or injury to the extent that this may be caused by something like turbulent blood flow, aka CCSVI.

Vitamin D expresses the gene that sets off the cell replication cycle in which a defective cell creates a copy of itself and then kills of the old, defective cell.

NZer1 wrote:If that infection and possible other infections that will cross the BBB to create the outcome of 'lesions and disability' the target for the treatment, then is the treatment anything to do with the symptoms? The outcome of symptoms 'can' be an incidental process (synergistic) that occurs because of a breached BBB, but not the pathogen itself or vice versa.

So is it the repaired immune system kill rate the makes a rapid recovery in the Prof H study or is it repair of the damage caused by a multi-pathogen infection within the BBB safe zone that has been entered or is it the repair of the BBB breaches or entry points?
Probably many more theories why the mice recover so quickly after a stunt of Calcitriol and Vit D?

Nigel,
The BBB is there to prevent almost everything from crossing into the CNS. Anything, blood cells, pathogens, parts of the body side immune system that crosses over will all cause a reaction of the CNS immune system which is different in many ways than what happens on the other side of the BBB.

But the principle injury that occurs in MS when the BBB is broken is that CD4+ T-cells cross over and tag myelin for destruction. The damage that results in the loss of a function nervous system in MS is demyelination and that happens because of the CD4+ T-cells.

So what happens with Prof Hayes' mice is that the single, high dose of calcitriol enters the CD4+ T-cells and initiates programmed cell death. It is the elimination of these T-cells with an affinity for myelin that halts the disease progression.

There is more to be done than just this. The immune system needs to start repairing the BBB. It also needs to eliminate any pathogens that are causing damage to the BBB. The results of her study show that after the single dose of calcitriol, quite modest doses of vitamin D3 are able to be converted to calcitriol and get on with fighting these other problems.

When her mice are given their dose of calcitriol, their little tails and hind legs are paralyzed. By the end of the study, they are wagging their tails and ambulating normally suggesting that recovery from damage to their nerves is possible when you attack the problem by supporting rather than suppressing the immune system.

I know I posted a reference to this study earlier, but since we are getting down to the level vitamin D metabolism, I thought I would note that this is a great resource to understand this intricate and complex system and its role in MS.

I've included a sample from the article so you can see the level of detail. You may need a little extra vitamin D to take all this in. :>)

Source URL: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3582312/
PMID: 23483715
DOI: 10.1177/1756285612473513
Journal Title: Therapeutic Advances in Neurological Disorders
Journal Date: March 2013
Journal Issue: 2
Journal Volume: 6
Journal First Page: 81
Abstract URL: http://www.ncbi.nlm.nih.gov/pmc/article ... t=abstract
Article Title: Contribution of vitamin D insufficiency to the pathogenesis of multiple sclerosis
Article Authors: Charles Pierrot-Deseilligny, Jean-Claude Souberbielle

The study by Sawcer and colleagues has shown the involvement in MS risk of two genes involved in vitamin D metabolism: CYP27B1, controlling 1-α-hydroxylase and therefore calcitriol synthesis, and CYP24A1, controlling calcitriol catabolism [Sawcer et al. 2011]. The genetic involvement of CYP27B1 in MS risk was also found in another GWAS considering a specific pathway in which eight genes within a module of 13 genes influenced by vitamin D were associated with MS [ANZgene and the Australia and New Zealand Multiple Sclerosis Genetics Consortium, 2009] and is now confirmed thanks to several other types of genetic research methods used to study patients with MS with vitamin D-dependent rickets [Torkildsen et al. 2008], with rare variants [Ramagopalan et al. 2011a] or with single nucleotide polymorphisms [Sundqvist et al. 2010; Simon et al. 2011]. However, it should be mentioned that no association was found between genes involved in vitamin D metabolism and MS risk in two other studies [Orton et al. 2011a; Smolders et al. 2011c], but these studies were performed with a much smaller sample than the one used by Sawcer and colleagues [Sawcer et al. 2011]. Furthermore, the genes encoding CYP27B1 and CYP24A1 could be epigenetically regulated [Kim et al. 2009; Novakovic et al. 2009], which may influence the vitamin D serum level and MS risk [Burrell et al. 2011]. Moreover, interactions between CYP27B1 and HLA-DRB1*15 may exist and influence the MS risk [Simon et al. 2011] (see below).

Ed the way I understand a gene task is that it is a switch that has an operator who can be remote and of another network.
If the genes are either on or off is to some degree irrelevant at the gene, the important factor is the switch mechanism.
For instance when switched on it directs an action one way and when off it directs either in another independent direction or not at all.
So the key player is the operator of the switch and not the switch.
So if diet is the operator of the switch it will require xyz product to move the switch, BUT if a pathogen is able to tamper with the switch then the resetting of the switch becomes more than a change due to supply of xyz product.

So in my understanding the gene involvement is one step removed from the task in most cases.

If in-sighting apoptosis is the fix with calcitriol and Vit D then it is the activation of the death process plus the clean up that allows the gene to switch back to the standard options rather than the mutated pathogen option. BUT what else is effected by the introduction of apoptosis??

And of course I might be wrong

Just noticed it's mid day and my porch light is flickering, so I'm off to have a sleep!

;)
Nigel

NZer1 wrote:Ed the way I understand a gene task is that it is a switch that has an operator who can be remote and of another network.
If the genes are either on or off is to some degree irrelevant at the gene, the important factor is the switch mechanism.
For instance when switched on it directs an action one way and when off it directs either in another independent direction or not at all.
So the key player is the operator of the switch and not the switch.
So if diet is the operator of the switch it will require xyz product to move the switch, BUT if a pathogen is able to tamper with the switch then the resetting of the switch becomes more than a change due to supply of xyz product.

So in my understanding the gene involvement is one step removed from the task in most cases.

If in-sighting apoptosis is the fix with calcitriol and Vit D then it is the activation of the death process plus the clean up that allows the gene to switch back to the standard options rather than the mutated pathogen option. BUT what else is effected by the introduction of apoptosis??

And of course I might be wrong

Just noticed it's mid day and my porch light is flickering, so I'm off to have a sleep!

;)
Nigel

Nigel,
We are WAY above my pay grade here, but I am familiar with how this process works in skin cells in a genetic disorder that runs in my family.

The regulatory hormone vitamin D causes the "expression" of the ATP2C1 gene at the beginning of the cell replication cycle. I am only familiar with this in skin cells and don't know if the same process is used in the brain. I know that ATP2C1 is found in the brain so it may be. And I further know that this gene is involved in this process in a way that is supposed to create tight junctions between cells which it doesn't. Another place where tight junctions between cells is critical of course is in the Blood Brain Barrier so it may well be a part of what we are looking at here.

A gene is "silent" until it is "expressed." In this case, when ATP2C1 is "expressed" it produces a protein which in turn creates a "transport receptor" on the wall of the Golgi Apparatus that allows ionized calcium, Ca++ (also notated as Ca2+). This transport receptor allows ionize calcium to pass through the wall of the Golgi Apparatus where the unbalanced Ca++ finds an ATP molecule with a friendly phosphorous atom willing to share electrons with the CA++, binding it to the ATP molecule which now contains calcium.

This new form of ATP +Ca then leaves the Golgi Apparatus, also through a transport receptor that allows it to do that. (Cells and all their internal compartments such as the Golgi, endoplasmic reticulum, nucleus have walls to keep things separated, unless a specific thing is needed which then passes through a special transport receptor which allows this to happen.)

Once the ATP + Ca molecule leaves the Golgi, it finds another transport receptor on the wall of the nucleus which allows it to enter the nucleus and set off the cell replication process which in turn triggers the destruction of the old cell one a new copy is made.

So, from this perspective, genes are "silent," sitting in the DNA waiting to be "expressed" and when "expressed" or activated, creates a protein that does something, in this case, make a transport receptor to allow ionized calcium to enter the Golgi.

So in your terminology, in this case, the "switch" is regulatory hormone vitamin D in the form of calcitriol and "on" and "off" are "silent," sitting in our DNA waiting to be activated, or "expressed."

I have looked at this in depth in terms of a genetic disorder in which one of the two copies of ATP2C1 is defective and results in a loss of the tight junction that exists in skin cells that prevent things from the outside entering the space between cells.

Although I know this gene is found in the brain and apparently used in cell replacement cycle, I don't know whether it is used solely for apoptosis outside the context of the creation of a new, replacement cell.

I also know that my description above at the level of individual ionized calcium atoms is a gross over simplification of what is going on. The whole process involves many more genes to create all the transport receptors involved in this process and likely do other things related to the cell replacement cycle.

Ed the reading I have done says that genes are naturally sitting in one mode and it is changed by the environment or the other cells it is linked to.

Is the process you are talking about with your disease a heme process of the ATP cycle, it sounds like it to me? If so it would be interlocked with secondary porphria issues which from memory have symptoms from a similar dysfunction in the heme cycle and your disease sounds similar to other life long skin diseases.

So any gene can be reset, the challenge is to find the keys that will do this.
Some studies now are saying that there are all types of signals that change gene expression and brain signals have been able to switch genes in experiments. So this is the classic work in process learning by Medicine although esoteric followers are already convinced.

So in regard to the cal/Vit D treatments it is another work and knowledge curve in process depending on the specialists that are involved.
Most of the processes in the body are long chain processes that have interplays with loops and going off at many stages and that is where we laymen are lost, imo!

An article I was sent this morning;
Abstract
Multiple sclerosis, MS (OMIM No. 126200), is a complex inflammatory disease that is characterized by lesions in the central nervous system. Both genes and other environmental factors influence disease susceptibility. One of the environmental factors that has been implicated in MS and autoimmune disease, such as type 1 diabetes, is vitamin D deficiency, in which patients have lower levels of 25-hydroxyvitamin D3 (25-OHD(3)) in blood than do controls. Previtamin D(3) is produced in the skin, and turned into 25-OHD(3) in the liver. In the kidney, skin and immune cells, 25-OHD(3) is turned into bioactive 1,25(OH)(2)D(3) by the enzyme coded by CYP27B1 (cytochrome P450 family 27 subfamily B peptide 1) on chromosome 12q13.1-3. 1,25(OH)(2)D(3) binds to the vitamin D receptor, expressed in T cells and antigen-presenting cells. 1,25(OH)(2)D(3) has a suppressive role in the adaptive immune system, decreasing T-cell and dendritic cell maturation, proliferation and differentiation, shifting the balance between T-helper 1 (Th1) and Th2 cells in favor of Th2 cells and increasing the suppressive function of regulatory T cells. Rs703842 in the 12q13-14 region was associated with MS in a recent study by the Australian and New Zealand Multiple Sclerosis Genetics Consortium (ANZgene). We show associations with three SNPs in this region in our Swedish materials (2158 cases, 1759 controls) rs4646536, rs10877012 and rs10877015 (P=0.01, 0.01 and 3.5 × 10(-3), respectively). We imputed rs703842 SNP and performed a joint analysis with the ANZgene results, reaching a significant association for rs703842 (P=5.1 × 10(-11); odds ratio 0.83; 95% confidence interval 0.79-0.88). Owing to its close association with 25-OHD(3), our results lend further support to the role of vitamin D in MS pathology.
http://www.ncbi.nlm.nih.gov/m/pubmed/20648053/

More on D3, relates to previous discussion on CSF levels.....................MarkW

Full paper - http://pmr.cuni.cz/file/5663/PMR2013A0019.pdf

Abstract:
25-hydroxyvitamin D3 Concentration
in Serum and Cerebrospinal Fluid
of Patients with Remitting-relapse
Multiple Sclerosis
Moghtaderi A.1, Tamadon G. H.2, Haghighi F.1
1Department of Neurology, Zahedan University of Medical Sciences, Zahedan, Iran;
2Department of Laboratory Medicine, Shiraz University of Medical Sciences,
Shiraz, Iran
Key words: Multiple sclerosis – CSF – Vitamin D – Plasma
Abstract: There is epidemiological, geographical and immunological evidence
suggesting that low environmental supplies of vitamin D3 may act as a risk
factor for developing multiple sclerosis (MS), possibly due to dysfunction in the
immunomodulatory properties of 25-hydroxyvitamin D3 (25-OH-D3) in the
brain. The objective of this study is to measure the serum and cerebrospinal fluid
(CSF) concentrations of 25-OH-D3 in MS patients during their relapsing phase.
52 patients with remitting-relapse and 58 patients with other non-inflammatory
diseases of central and peripheral nervous system were entered into the study.
Patients in both groups were admitted for the first time to do diagnostic
procedures and they were not on any other treatment for neurological disorders.
The means and medians for serum levels of 25-OH-D3 in MS patients and control
group were 10.64 ± 9.2 ng/ml (median: 9.6 ng/ml) and 13.23 ± 17.56 ng/ml (median:
11.90 ng/ml), respectively (p=0.328). CSF concentrations for the same values were
2.02 ± 1.94 ng/ml (median: 0.23 ng/ml) and 3.28 ± 2.96 (median: 0.29 ng/ml),
respectively (p=0.242). The differences between calculated numbers of serum/CSF
ratios were not statistically significant too. The serum and CSF concentrations
of 25-OH-D3 in MS group were lower than the control counterpart without any
statistical difference and the authors did not find any influence of serum 25-OH-D3
concentration on the CSF concentration based on the non-significant statistical
difference between the serum/CSF ratios in two study groups of MS patients
and control cases.
Mailing Address: Assoc. Prof. Ali Moghtaderi, MD., Department of Neurology,